Multi-comparative Thermal Proteome Profiling Uncovers New O‑GlcNAc Proteins in a System-wide Method

Among
diverse protein post-translational modifications, O-GlcNAcylation,
a simple but essential monosaccharide modification, plays crucial
roles in cellular processes and is closely related to various diseases.
Despite its ubiquity in cells, properties of low stoichiometry and
reversibility are hard nuts to crack in system-wide research of O-GlcNAc.
Herein, we developed a novel method employing multi-comparative thermal
proteome profiling for O-GlcNAc transferase (OGT) substrate discovery.
Melting curves of proteins under different treatments were profiled
and compared with high reproducibility and consistency. Consequently,
proteins with significantly shifted stabilities caused by OGT and
uridine-5′-diphosphate N-acetylglucosamine
were screened out from which new O-GlcNAcylated proteins were uncovered.